CN101906397A - Fermentation process of trichoderma transformant of anti-benzimidazole type chemical fungicide - Google Patents
Fermentation process of trichoderma transformant of anti-benzimidazole type chemical fungicide Download PDFInfo
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- CN101906397A CN101906397A CN2010101890303A CN201010189030A CN101906397A CN 101906397 A CN101906397 A CN 101906397A CN 2010101890303 A CN2010101890303 A CN 2010101890303A CN 201010189030 A CN201010189030 A CN 201010189030A CN 101906397 A CN101906397 A CN 101906397A
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Abstract
The invention belongs to the technical field of microbial pesticides. The invention relates to a fermentation process of a trichoderma transformant of an anti-benzimidazole type chemical fungicide, which comprises the following steps: taking a trichoderma harzianum transformant TS1 as a strain; taking maize stem dust and wheat bran after screening as a solid culture medium, adding water, mixing, and carrying out sterilization at high temperature and high pressure; and inoculating the transformant strain, carrying out fermentation in a conical flask and preparing the strain for amplification culture. The specific steps are as follows: screening the maize stem dust and the wheat bran as raw materials, adding the water, mixing, carrying out the sterilization at high temperature and high pressure, inoculating the mixture of the solid culture medium and the strain, transferring into a sterile fermentation bag, carrying out ventilation fermentation, preparing a trichoderma conidiophore fermentation culture and carrying out the fermentation for 5 days. A trichoderma conidiophore microbial novel pesticide and a formulation produced by fermentation according to the invention can lead the effects of preventing and treating phytophthora (potato late blight, root rot of different fruit tress and the like), fusarium (tomato wilt disease, the root rot of different plants and the like) and other soil-borne diseases to be more than 80%.
Description
Technical field
The invention belongs to the microbial pesticide technical field, particularly a kind of biological control agents of anti-benzimidazole type chemical fungicide--the zymotechnique of wooden mould transformant.
Background technology
Plant diseases has become the important factor of the human agriculture forest and husbandry production output of influence, and disaster often causes the underproduction even total crop failure when taking place, and brings about great losses to agriculture forest and husbandry.Utilizing the chemical pesticide control disease pest and weed is an important means that improves crop yield in China, but increase year by year along with its usage quantity, chemical pesticide because of its inherent toxicity and residual severe contamination human environment of depending on for existence, destroyed the eubiosis, cause great threat to human beings'health, and use chemical pesticide to make phytopathogen produce resistance for a long time to sterilant, directly influenced the prevention effect of chemical pesticide in the pest management system.Benzimidazole germicide has outstanding interior absorption and very high sterilization effect, be widely used, so its resistance problem is particularly serious, and the harm that is caused is also maximum.
Biological pesticide has the specificity of height, prevents and treats disease efficient height, and the natural enemy that does not does not kill and wound insect is also nontoxic to people and animals, and phytopathogen is developed immunity to drugs, and easily degraded, and noresidue can strengthen the disease resistance of plant, the stimulating plant growth.But the biological pesticide prevention effect is slower, uses biological pesticide to control crop pest separately, often is subjected to the interference of factors such as envrionment conditions, causes the field control effect instability, makes agriculture production bear very big risk.Therefore, the compound use of chemical pesticide and biological pesticide has become a new way of present control of plant disease.Trichoderma harziarum is a kind of good control of plant disease fungi, because to the high susceptibility of benzimidazole germicide, has limited the compound use of itself and sterilant.The Yang Qian of Harbin Institute of Technology etc. adopt engineered method, utilize the benzimidazole germicide resistant gene to transform Trichoderma, obtained can genetic stability, have a transformant T to the high resistance level of benzimidazole germicide
S1(Min Li, Qian Yang.Isolation andcharacterization of a β-tubulin gene from Trichodermaharzianum.Biochemical genetics, 2007,45:529-534.) the benzimidazole germicide resistant gene is imported in the bacterial strain, can be applied to the integrated control of Plant diseases with benzimidazole germicide simultaneously, help the control of disease and the usage quantity of minimizing chemical agent, reduce environmental pollution effectively.
But, will developing exactly of being badly in need of at present solving is efficient, economical and possess the strain fermentation process of commercial production conditions, and the formulation and the application process of definite bacterium medicine compound disinfectant, to improve the adaptability and the biological and ecological methods to prevent plant disease, pests, and erosion effect of bacterial strain, to solve the active demand in the agriculture production of current China.
Summary of the invention
The present invention seeks to prepare a kind of efficient, economical, the zymotechnique of trichoderma transformant that possesses the anti-benzimidazole type chemical fungicide of commercial production conditions, determine the formulation and the application process of compound disinfectant simultaneously, to improve the adaptability and the biological and ecological methods to prevent plant disease, pests, and erosion effect of bacterial strain.
The zymotechnique preparation method of the trichoderma transformant of anti-benzimidazole type chemical fungicide of the present invention is as follows:
Step 1:
With trichoderma harziarum transformant T
S1Be bacterial strain; With the corn stalk powder after the meticulous screening, Testa Tritici, add certain proportion water and mix, make the solid culture based raw material, autoclave sterilization; Inoculation transformant t bacteria
S1, adopting Erlenmeyer flask fermentative preparation enlarged culturing bacterial classification, cultured continuously is 5 days in insulation can.
Step 2:
With corn stalk powder, Testa Tritici after the screening is culture medium raw material, adds certain proportion water and mixes autoclave sterilization, inoculation solid medium and bacterial classification mixture, change aseptic plastics fermentation bag over to, ventilating fermentation prepares the trichoderma conidium fermenting culture, continuously ferments 5 days promptly.
Substratum after the fermentation and fungi are together made the pesticide formulation and render in the farmland, be fungi initial habitat is provided, helping it survives, do not produce the residue contamination environment again, can incorporate edatope again, realize straw-returning, increase soil fertility, achieve many things at one stroke, when solving problem of environmental pollution, realized the comprehensive utilization of resource.
The present invention is a bacterial strain with the trichoderma transformant of anti-benzimidazole type chemical fungicide, the fermentative production trichoderma conidium is also made formulation, developing the novel transformant with the Trichoderma anti-benzimidazole type chemical fungicide is core novel microorganism agricultural chemicals, and this novel agrochemical reaches more than 80% soil-borne disease prevention effect such as phytophthoras (late blight of potato, different fruit tree root rot etc.), sickle-like bacteria (that tomato withers is here sick, the root rot of different plants etc.).
Embodiment
The preparation method's of wooden removing mildew of the present invention concrete implementation step is as follows:
(1) with the corn stalk powder 2.5g after the screening of 60 mesh sieves, Testa Tritici 5g is the solid culture based raw material, adds 10ml water, mixes autoclave sterilization, cooling; The transformant trichoderma harziarum T of inoculation anti-benzimidazole type chemical fungicide
S1Bacterial classification, sterilization ring picking 4-6 time stirs several times in the 250ml Erlenmeyer flask, seals, and cultivates in 28 ℃ of incubators, rocked once every 24 hours, cultured continuously 5 days, stand-by as enlarged culturing with bacterial classification.
(2) with corn stalk powder 5g, Testa Tritici 10g after the screening of 40 mesh sieves, add entry 20ml and mix, be culture medium raw material, autoclave sterilization, cooling; The enlarged culturing of inoculation step (1) gained solid medium and bacterial classification mixture 0.5g, change aseptic plastics fermentation bag over to, pass to aseptic wind, ventilating fermentation prepares the trichoderma conidium fermenting culture in 28 ℃ incubator, stirred once in 24 hours, continuously fermenting promptly obtained the culture of trichoderma conidium in 5 days.
Production application
Field experiment shows: this microorganism novel agrochemical reaches more than 80% soil-borne disease prevention effect such as phytophthoras (late blight of potato, different fruit tree root rot etc.), sickle-like bacteria (that tomato withers is here sick, the root rot of different plants etc.).This microorganism novel agrochemical can not produce 3R problem---the resistance of chemical insecticide, wildness and pesticide residue again.Substratum after the fermentation and fungi are together made the pesticide formulation and rendered in the farmland, be fungi initial habitat is provided, helping it survives, can incorporate edatope again, do not produce the residue contamination environment, realize straw-returning, increase soil fertility,-lift get more, when solving problem of environmental pollution, realized the comprehensive utilization of resource.
Claims (6)
1. the zymotechnique of the trichoderma transformant of an anti-benzimidazole type chemical fungicide, it is characterized in that: the preparation method is as follows for this zymotechnique
Step 1:
With trichoderma harziarum transformant T
S1Be bacterial strain; With the corn stalk powder after the meticulous screening, Testa Tritici, add certain proportion water and mix, make the solid culture based raw material, autoclave sterilization; Inoculation transformant t bacteria
S1, adopting Erlenmeyer flask fermentative preparation enlarged culturing bacterial classification, cultured continuously is 5 days in insulation can;
Step 2:
With corn stalk powder, Testa Tritici after the screening is culture medium raw material, adds certain proportion water and mixes autoclave sterilization; Inoculation solid medium and bacterial classification mixture change aseptic plastics fermentation bag over to, and ventilating fermentation prepares the trichoderma conidium fermenting culture, continuously ferments 5 days promptly.
2. the zymotechnique of trichoderma transformant according to claim 1, it is characterized in that: the described solid culture based raw material of step 1, promptly with the corn stalk powder 2.5g after the screening of 60 mesh sieves, Testa Tritici 5g, be the solid culture based raw material, add 10ml water, mix, autoclave sterilization is made after the cooling.
3. the zymotechnique of trichoderma transformant according to claim 1 is characterized in that: the described inoculation transformant of step 1 t bacteria
S1, be about to transformant trichoderma harziarum T
S1Bacterial classification, sterilization ring picking 4-6 time stirred several times in the 250ml Erlenmeyer flask, seals, and cultivates in 28 ℃ of incubators, rocked once cultured continuously 5 days every 24 hours.
4. the zymotechnique of trichoderma transformant according to claim 1, it is characterized in that: the described culture medium raw material of step 2 is that corn stalk powder 5g, Testa Tritici 10g with after the screening of 40 mesh sieves add entry 20ml and mix, autoclave sterilization is made after the cooling.
5. the zymotechnique of trichoderma transformant according to claim 1, it is characterized in that: described inoculation solid medium of step 2 and bacterial classification mixture, be about to the enlarged culturing solid medium and the bacterial classification mixture 0.5g of step 1 gained, change aseptic plastics fermentation bag over to, pass to aseptic wind, ventilating fermentation prepares the trichoderma conidium fermenting culture in 28 ℃ incubator, stirs once in 24 hours, and continuously fermenting promptly obtained the culture of trichoderma conidium in 5 days.
6. the zymotechnique of the trichoderma transformant of an anti-benzimidazole type chemical fungicide is characterized in that: the substratum after the fermentation of the present invention and fungi together can be made the pesticide formulation and render in the farmland.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2010101890303A CN101906397A (en) | 2010-06-02 | 2010-06-02 | Fermentation process of trichoderma transformant of anti-benzimidazole type chemical fungicide |
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| CN2010101890303A CN101906397A (en) | 2010-06-02 | 2010-06-02 | Fermentation process of trichoderma transformant of anti-benzimidazole type chemical fungicide |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102732434A (en) * | 2012-06-18 | 2012-10-17 | 中国农业大学 | Trichoderma harzianum strain, and its application in control of potato late blight |
-
2010
- 2010-06-02 CN CN2010101890303A patent/CN101906397A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102732434A (en) * | 2012-06-18 | 2012-10-17 | 中国农业大学 | Trichoderma harzianum strain, and its application in control of potato late blight |
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Open date: 20101208 |